Device

Part:BBa_K1150042

Designed by: Ilona Blank   Group: iGEM13_Freiburg   (2013-09-17)

uniCAS RNaimer to Bla Target 4

RNaimer (Bla target 4)
Function Targeting VEGF4 with Cas9
Use in Mammalian cells
RFC standard RFC 25
Backbone pSB1C3
Submitted by [http://2013.igem.org/Team:Freiburg Freiburg 2013]

This device contains loci coding for small RNAs that are able to target the catalytically-inactive, engineered dCas9 protein to the Bla locus target 4 . This RNaimer is bulid of the uniCAS RNAimer and contains the DNA sequence coding for a crRNA that binds complementary to Bla 4 target.


Usage and Biology

Freiburg 2013 used this plasmid to test the efficiency of target Bla4 by activating or repressing a SEAP reporter gene.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 224
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 216


Functional Parameters



Functional Parameters: Austin_UTexas

Burden Imposed by this Part:

Burden Value: 7.8 ± 2.1%

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This part exhibited a significant burden. Users should be aware that BioBricks with a burden of >20-30% may be susceptible to mutating to become less functional or nonfunctional as an evolutionary consequence of this fitness cost. This risk increases as they used for more bacterial cell divisions or in larger cultures. Users should be especially careful when combining multiple burdensome parts, as plasmids with a total burden of >40% are expected to mutate so quickly that they become unclonable. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.

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Categories
Parameters
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